加味丹参抗康口腔黏膜损伤的作用：网络药理学和联合超高效液相色谱（UPLC）和质谱（MS）。

PURPOSE: Arecoline is one of the main toxic components of arecoline to cause oral mucosal lesions or canceration, which seriously affects the survival and life quality of patients. This study analyzed the mechanism of Jiawei Danxuan Koukang (JDK) in alleviating arecoline induced oral mucosal lesions, to provide new insights for the treatment of oral submucosal fibrosis (OSF) or cancerosis. METHODS: Metabolomics was applied to analyze the composition of JDK and serum metabolites. The active ingredients of JDK were analyzed by the combined ultra-high performance liquid chromatography and mass spectrometry. The target network of JDK, metabolites and OSF was analyzed by network pharmacology, and molecular docking. Oral mucosal lesions and fibrosis were analyzed by HE and Masson staining. Cell differentiation, proliferation and apoptosis were detected. The expressions of α-SMA, Collagen I, Vimentin, Snail, E-cadherin, AR and NOTCH1 were detected by Western blot. RESULTS: Arecoline induced the gradual atrophy and thinning of rat oral mucosal, collagen accumulation, the increase expressions of fibrosis-related proteins and Th17/Treg ratio. JDK inhibited arecoline-induced oral mucosal lesions and inflammatory infiltration. Arecoline induced changes of serum metabolites in Aminoacyl-tRNA biosynthesis, Alanine, aspartate and glutamate metabolism and Arginine biosynthesis pathways, which were reversed by M-JDK. Quercetin and AR were the active ingredients and key targets of JDK, metabolites and OSF interaction. Arecoline promoted the expression of AR protein, and the proliferation of oral fibroblasts. Quercetin inhibited the effect of arecoline on oral fibroblasts, but was reversed by AR overexpression. Arecoline induced NOTCH1 expression in CAL27 and SCC-25 cells, and promoted cell proliferation, but was reversed by M-JDK or quercetin. CONCLUSION: JDK improved the arecoline-induced OSF and serum metabolite functional pathway. Quercetin targeted AR protein to improve arecoline-induced OSF. JDK and quercetin inhibited arecoline-induced NOTCH1 protein expression in CAL27 and SCC-25 cells to play an anti-oral cancer role.

目的：槟榔碱是导致口腔黏膜病变或癌变的主要毒性成分之一，严重影响患者的生存和生活质量。本研究分析了加味丹玄抗康（JDK）缓解槟榔碱诱导的口腔黏膜病变的机制，为口腔黏膜下纤维化（OSF）或癌变的治疗提供新的思路。

方法：采用代谢组学分析 JDK 及血清代谢物的组成，采用超高效液相色谱与质谱联用分析 JDK 的活性成分，采用网络药理学和分子对接分析 JDK、代谢物与 OSF 的靶标网络，通过 HE 和 Masson 染色分析口腔黏膜病变和纤维化，检测细胞分化、增殖和凋亡，采用 Western blot 检测α-SMA、Collagen I、Vimentin、Snail、E-cadherin、AR 和 NOTCH1 的表达。

结果：槟榔碱诱导大鼠口腔黏膜逐渐萎缩变薄，胶原堆积，纤维化相关蛋白表达增加，Th17/Treg 比例升高。JDK 抑制槟榔碱诱导的口腔黏膜病变和炎症浸润。槟榔碱诱导 Aminoacyl-tRNA biosynthesis、Alanine, aspartate and glutamate metabolism 和 Arginine biosynthesis 通路的血清代谢物发生变化，M-JDK 可逆转这些变化。槲皮素和 AR 是 JDK、代谢物与 OSF 相互作用的活性成分和关键靶点。槟榔碱促进 AR 蛋白表达和口腔成纤维细胞增殖，槲皮素抑制槟榔碱对口腔成纤维细胞的作用，但被 AR 过表达所逆转。槟榔碱诱导 CAL27 和 SCC-25 细胞中 NOTCH1 表达，并促进细胞增殖，但被 M-JDK 或槲皮素所逆转。

结论：JDK 改善了槟榔碱诱导的 OSF 和血清代谢物功能途径。槲皮素靶向 AR 蛋白，改善槟榔碱诱导的 OSF。JDK 和槲皮素抑制 CAL27 和 SCC-25 细胞中槟榔碱诱导的 NOTCH1 蛋白表达，发挥抗口腔癌作用。